Immune cells in dorsal root ganglia are associated with pruritus in a mouse model of allergic contact dermatitis and co-culture study

The interaction between the neuroimmune system plays a crucial role in itch sensation, yet most research has focused on immune cells within the skin. Our study seeks to explore the presence and functions of immune cells within the dorsal root ganglia (DRG) in the context of allergic contact dermatitis (ACD). Immunofluorescence and histological staining techniques were employed to identify immune cells, including T-cells, basophils, mast cells, and dendritic cells (DCs), within the DRG of BALB/c mice sensitized and challenged with toluene diisocyanate (TDI). Our findings revealed an increase in mast cells and DCs within the DRG under ACD condition. Additionally, when DRG neurons were cultured with mast cells, a higher proportion of neurons exhibited responses to non-histaminergic pruritogens compared to neurons cultured alone. This suggests that mast cells may contribute to heightened sensitivity to non-histaminergic pruritogens. Furthermore, we conducted transcriptomic analysis of DCs within the DRG using RNA sequencing, followed by pathway enrichment analysis. Our analysis revealed that sorted DCs are implicated in immune responses, inflammation, and itch, with notable upregulation of Cathepsin S (Ctss) and sphingosine-1-phosphate (S1P) phosphatase 2 (Sgpp2). Subsequent functional experiments targeting CTSS in co-culture studies validated suppressed response to pruritogen and agonists of TRPA1 and TRPV1, indicating a potential role in peripheral sensitization. Additionally, the co-culture study indicated that the neuroimmune interaction between DCs and DRG neurons might involve S1P metabolism and S1P receptor signaling. In conclusion, targeting DCs and exploring the non-histaminergic functions of mast cells within the DRG, holds promise as novel targets for treating pruritus.